PLASMA PROTEINS.

Albumin.
• Albumin is the most abundant circulating protein found in plasma. 

• Its molecular weight is 66.5 kilodaltons (kDa).

• It consists of 585 amino acids.

• Albumin is also a colloid fluid administered to patients in need of fluid

resuscitation, especially in the setting of trauma (i.e. hypovolemic shock).

• Resuscitation is the process of correcting physiological disorders (such as lack

of breathing or heartbeat) in an acutely ill patient

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 Clinical applications of Albumin.

• Albumin is therapeutically useful.

• It is used to treat or prevent shock following serious injury, bleeding,

surgery, or burns by increasing the volume of blood plasma. This
medicine can also replace low blood protein.

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• Kidney diseases

• chronic kidney disease(CKD leads to lower the albumin levels. Kidney

disease can result in albumin spilling into the urine, in cases like,
nephrotic syndrome, certain inflammatory conditions of urinary tract.

•  Microalbuminuria, is a clinically important for predicting the future risk

of renal diseases.

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• Liver diseases: Lower levels of albumin are also seen in cases of liver
diseases, such as hepatitis (an inflammation of the liver, often due to viral
infection).

•  Liver cirrhosis (scarring of liver tissue due to chronic hepatitis or

excessive alcohol use), jaundice , and others.

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• Crohn’s disease: A persistent and damaging inflammation of the
gastrointestinal tract, Crohn’s disease leads to difficulties absorbing nutrients.
In these cases, albumin levels are lower than normal. 

• Whipple disease: This rare bacterial infection affects joints and the digestive
system, impairing the body’s ability to digest food. In these cases, albumin
levels will be lower than normal.
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• Malnutrition: Decreased levels of albumin in the blood may also be
signs of malnutrition.

• Dehydration: Insufficient water in the body will cause albumin levels

to be elevated.

• Medications and therapies: Some medications, including insulin,

steroids will raise levels; whereas others, such as birth control pills,
will lower them.
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• Albumin test may be investigated in one or more of the following
symptoms are present:

• unexpected weight loss

• swelling around the stomach, eyes, or legs

• jaundice, causing yellowing of the skin and eyes

• unexplained fatigue
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• A review has indicated that Surgical site infection (SSI) is an unfortunately
common postoperative complication encountered in the field of
orthopaedic surgery (e.g. orthopaedic spine surgery, elective total joint
replacement, orthopaedic trauma cases). 

• Serum albumin <3.5 g/dL has been demonstrated to confer an increased

risk of SSI. 

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Hypoalbuminemia Hyperalbuminemia
Nephrotic syndrome Dehydration
Burns High protein diet
Blood loss False value due to prolonged tourniquet
Malignancies
Inflammatory process
Liver diseases
Decreased protein intake
Ascites
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 Globulins.
• The globulins are one of the major classifications of proteins,  that have
higher molecular weights than albumins.

• Globulins are proteins in the blood that help to protect the body.

•  They are insoluble in pure water but dissolve in dilute salt solutions. 

• They are categorized in to alpha, beta, and gamma globulins.

• Globulins can be distinguished from one another using serum protein

electrophoresis.
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• Alpha and beta globulins are transport proteins.

• Gamma globulins have a vital role in natural and acquired immunity

to infection

• Globulin concentration is determined by subtraction: [total protein] −

[albumin] = [globulin]; therefore errors in those values can lead to
erroneous globulin values.

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 Globulin : Normal range : 2.5-3.5 g/dl

• Low Globulin Levels. If the globulin levels fall below the normal range it can
be a sign of several serious health conditions.

• Renal disease, hepatic dysfunction, celiac disease, inflammatory bowel disease

(IBD) and acute haemolytic anaemia can cause the globulin levels to drop.

• This is also a sign that proteins taken in by the digestive system are not being
broken down or absorbed properly.

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• High globulin levels. Those with high globulin levels may be suffering
from leukemia or other bone marrow disorders, autoimmunity diseases,
chronic inflammatory diseases such as syphilis, liver disease, Rheumatoid
arthritis, kidney disease or a chronic viral or bacterial infection.

• Further testing will be necessary to determine which of these disorders is

causing the globulin levels to rise so adequate treatment can be
administered.
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 α1- antitrypsin.
• α1- antitrypsin, also referred as α1- proteinase, made by the liver.

• It protects the body's tissues from being damaged by infection-fighting

agents released by its immune system.

• Its mol.wt is 54,000.

• Its normal range is around 200 mg/ dl.

• Alpha1-antitrypsin deficiency (AATD, AAT deficiency) is an inherited

condition that increases the risk of lung and liver disease.
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• Its main function is to protect the lungs.

• If these proteins are malformed or deficient, the impact is

predisposition for obstructive pulmonary disease and liver disease.

• α1-Antitrypsin deficiency has been implicated in the diseases, namely,

emphysema ( to inflate, explaining abnormal distension of lungs by air
).
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• This is associated with lung infections (e.g. pneumonia).

• And increase in the activity of macrophages to release elastase that

damages lung tissues.

• In the normal circumstances, elastase activity is inhibited by alpha-1-AT.

• Elastase is an enzyme found in the pancreas.

• Elastase helps break down fats, proteins, and carbohydrates. It's a key part
of the digestive process. 
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• Emphysema is more commonly associated with heavy smoking and
the situation becomes worse in persons with alpha-1-AT deficiency.

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 α-2-Macroglobulin
• It is a protein,( mol. wt-= 8,00, 000) produced by the liver.

• It acts as an antiprotease and is able to inactivate an enormous variety of

proteinases. ( normal range =  178.8 ± 8.5 mg/ml )

• It functions as an inhibitor of coagulation by inhibiting thrombin.

• Its concentration is elevated in nephrotic syndrome.

•  This is due to the fact that majority of the low molecular weight proteins are lost in
urine (proteinuria) in this disorder. ( High mol.wt, α-2-Macroglobulin gets retained in
the body. 22
 Albumin to globulin ratio.
• The normal A/G ratio is 0.8-2.0.

• Serum albumin/globulin ratio (AGR) can predict the prognosis of various

diseases, including cancers, chronic kidney disease, heart failure,
nutritional problems.

• Total protein ( value) – Albumin ( value ) = Globulin.

• 7.6 – 3.9 = 3.7. A / G ratio…..3.9 / 3.7 = 1.1

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• Help full in assessing nutritional status of hospitalized patients, especially
geriatric patients.

• Decrease in albumin (without decrease in globulins).

• Decreased production.; Severe liver disease .

• Inflammation .;

• Protein-losing nephropathy (PLN);

• Neoplasia.;
• Multiple myeloma .

• Increase catabolism.

• Thyroid dysfunction (related to overproduction of albumin binding

thyroid hormones).

• Congestive heart failure.

• Interfering factors ( drugs )

 Albumin Globulin

A single protein, A group of proteins.

Normal range 3.5- 5.0 gm /dl. Normal range 2.5- 4.5 gm /dl

Molecular weight: 66.5 kDa Molecular weight, comparatively higher.

Important in increasing the Can be the enzymes, immunoglobulins.

osmotic pressure of the blood.

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Protein separation.

• To study proteins, it is important to first identify and isolate them.

• it needs to carry high-throughput protein purification.

• First, individual proteins are isolated, then they are separated based
on molecular weight, solubility, charge, and specific binding affinity. 

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 salting out
• Salting out is a purification method that utilizes the reduced solubility of
proteins.

• Salting out occurs in aqueous solutions of high ionic strength that reduce the
molecule's solubility causing certain proteins to precipitate.

• Salting out is an effective means for initial molecule purification, but lacks the
ability for precise isolation of a specific protein.
• The salt commonly used is ammonium sulfate because:

• Its large solubility in water.

• Its relative freedom from temperature effects.

• The salt concentration at which a protein precipitates differs from one protein
to another.

• Hence, salting out can be used to fractionate proteins. For example, 0.8 M
ammonium sulfate precipitates fibrinogen, a blood-clotting protein, whereas a
concentration of 2.4 M is needed to precipitate serum albumin.
Protein electrophoresis
• Protein electrophoresis is a test that measures specific proteins in the blood.
The test separates proteins in the blood based on their electrical charge,
size, and shape.

• Electrophoresis had been widely used in clinical medicine for aiding in

diagnosis of various clinical conditions like acute and chronic inflammations,,
nephropathy, liver diseases, etc. 
• The separation of proteins by electrophoresis is based on the fact that charged
molecules usually migrate through a matrix/medium upon application of an
electrical field.

• The protein move in the electric field is dependent on certain factors like, strength
of the electric field, temperature of the system, pH of the ions, concentration of
buffer etc.

• Smaller proteins usually migrate faster, and larger proteins take a longer time. This
physical property of proteins is exploited for its separation by employing the
electrophoretic technique.
• Each of these protein fractions is distinct and at specific concentrations.

• The patterns typically seen in certain conditions and diseases can help with
diagnosis. 

•  
• The Cellulose acetate or agarose are used to separate proteins on the
basis of molecular charge.

• At one end, serum sample is applied and  a strong electrical current then
is passed through the medium for a set time period.

• During this time the proteins migrate from the application point
toward the positively charged anode for varying distances related
to their charge, size, and shape.
• After a set period, the gel with the separated proteins is removed and
immersed in a solution which stains the protein. ( bromophenol- blue)

•  After drying, the gel can be inspected visually for the distribution of bands.

• Albumin is the most abundant single protein in serum and forms a single
distinct band in the gel.

• The globulins are further separated into as many as other bands,

depending on the species and the underlying disease processes.
• The gel is then placed in an instrument, called a densitometer, which
determines the amount of protein in each band by its absorbance of
light.

• The densitometer translates this information into a series of peaks

called an electrophoretogram.
Densitometer.
 Chromatography
• Chromatography is the analytical technique used to separate mixture of a
proteins on the basis of size, affinity or ionic interaction.

• So that the individual components can be thoroughly analyzed. There are

many types of chromatography.

• e.g., liquid chromatography, gas chromatography, ion-exchange

chromatography, affinity chromatography.

• but all of these employ the same basic principles.

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• Sample to be examined called solute or analyte allowed to interact with
two immiscible phases-mobile and stationary phase.

• These two phases could be solid and liquid.

• The mobile phase may be either a liquid or a gas, while the stationary
phase is either a solid or a liquid.
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